Stable Isotope Tracing Uncovers Reduced γ/β-ATP Turnover and Metabolic Flux Through Mitochondrial-Linked Phosphotransfer Circuits in Aggressive Breast Cancer Cells

Aleksandr Klepinin; Sten Miller; Indrek Reile; Marju Puurand; Egle Rebane-Klemm; Ljudmila Klepinina; Heiki Vija; Song Zhang; Andre Terzic; Petras Dzeja; Tuuli Kaambre

doi:10.3389/fonc.2022.892195

Stable Isotope Tracing Uncovers Reduced γ/β-ATP Turnover and Metabolic Flux Through Mitochondrial-Linked Phosphotransfer Circuits in Aggressive Breast Cancer Cells

Aleksandr Klepinin, Sten Miller, Indrek Reile, Marju Puurand, Egle Rebane-Klemm, Ljudmila Klepinina, Heiki Vija, Song Zhang, Andre Terzic, Petras Dzeja, Tuuli Kaambre

Cardiovascular Medicine

Research output: Contribution to journal › Article › peer-review

Abstract

Changes in dynamics of ATP γ- and β-phosphoryl turnover and metabolic flux through phosphotransfer pathways in cancer cells are still unknown. Using ¹⁸O phosphometabolite tagging technology, we have discovered phosphotransfer dynamics in three breast cancer cell lines: MCF7 (non-aggressive), MDA-MB-231 (aggressive), and MCF10A (control). Contrary to high intracellular ATP levels, the ¹⁸O labeling method revealed a decreased γ- and β-ATP turnover in both breast cancer cells, compared to control. Lower β-ATP[¹⁸O] turnover indicates decreased adenylate kinase (AK) flux. Aggressive cancer cells had also reduced fluxes through hexokinase (HK) G-6-P[¹⁸O], creatine kinase (CK) [CrP[¹⁸O], and mitochondrial G-3-P[¹⁸O] substrate shuttle. Decreased CK metabolic flux was linked to the downregulation of mitochondrial MTCK1A in breast cancer cells. Despite the decreased overall phosphoryl flux, overexpression of HK2, AK2, and AK6 isoforms within cell compartments could promote aggressive breast cancer growth.

Original language	English (US)
Article number	892195
Journal	Frontiers in Oncology
Volume	12
DOIs	https://doi.org/10.3389/fonc.2022.892195
State	Published - May 31 2022

Keywords

18 O stable isotope labeling technology
adenylate kinase
creatine kinase
glycolysis
oxidative phosphorylation
phosphotransfer network
triple-negative breast cancer
γ-and β-ATP phosphoryl turnover

ASJC Scopus subject areas

Oncology
Cancer Research

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10.3389/fonc.2022.892195

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Klepinin, A., Miller, S., Reile, I., Puurand, M., Rebane-Klemm, E., Klepinina, L., Vija, H., Zhang, S., Terzic, A., Dzeja, P., & Kaambre, T. (2022). Stable Isotope Tracing Uncovers Reduced γ/β-ATP Turnover and Metabolic Flux Through Mitochondrial-Linked Phosphotransfer Circuits in Aggressive Breast Cancer Cells. Frontiers in Oncology, 12, Article 892195. https://doi.org/10.3389/fonc.2022.892195

Klepinin, A, Miller, S, Reile, I, Puurand, M, Rebane-Klemm, E, Klepinina, L, Vija, H, Zhang, S, Terzic, A, Dzeja, P & Kaambre, T 2022, 'Stable Isotope Tracing Uncovers Reduced γ/β-ATP Turnover and Metabolic Flux Through Mitochondrial-Linked Phosphotransfer Circuits in Aggressive Breast Cancer Cells', Frontiers in Oncology, vol. 12, 892195. https://doi.org/10.3389/fonc.2022.892195

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abstract = "Changes in dynamics of ATP γ- and β-phosphoryl turnover and metabolic flux through phosphotransfer pathways in cancer cells are still unknown. Using 18O phosphometabolite tagging technology, we have discovered phosphotransfer dynamics in three breast cancer cell lines: MCF7 (non-aggressive), MDA-MB-231 (aggressive), and MCF10A (control). Contrary to high intracellular ATP levels, the 18O labeling method revealed a decreased γ- and β-ATP turnover in both breast cancer cells, compared to control. Lower β-ATP[18O] turnover indicates decreased adenylate kinase (AK) flux. Aggressive cancer cells had also reduced fluxes through hexokinase (HK) G-6-P[18O], creatine kinase (CK) [CrP[18O], and mitochondrial G-3-P[18O] substrate shuttle. Decreased CK metabolic flux was linked to the downregulation of mitochondrial MTCK1A in breast cancer cells. Despite the decreased overall phosphoryl flux, overexpression of HK2, AK2, and AK6 isoforms within cell compartments could promote aggressive breast cancer growth.",

keywords = "18 O stable isotope labeling technology, adenylate kinase, creatine kinase, glycolysis, oxidative phosphorylation, phosphotransfer network, triple-negative breast cancer, γ-and β-ATP phosphoryl turnover",

author = "Aleksandr Klepinin and Sten Miller and Indrek Reile and Marju Puurand and Egle Rebane-Klemm and Ljudmila Klepinina and Heiki Vija and Song Zhang and Andre Terzic and Petras Dzeja and Tuuli Kaambre",

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doi = "10.3389/fonc.2022.892195",

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AU - Klepinin, Aleksandr

AU - Miller, Sten

AU - Reile, Indrek

AU - Puurand, Marju

AU - Rebane-Klemm, Egle

AU - Klepinina, Ljudmila

AU - Vija, Heiki

AU - Zhang, Song

AU - Terzic, Andre

AU - Dzeja, Petras

AU - Kaambre, Tuuli

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AB - Changes in dynamics of ATP γ- and β-phosphoryl turnover and metabolic flux through phosphotransfer pathways in cancer cells are still unknown. Using 18O phosphometabolite tagging technology, we have discovered phosphotransfer dynamics in three breast cancer cell lines: MCF7 (non-aggressive), MDA-MB-231 (aggressive), and MCF10A (control). Contrary to high intracellular ATP levels, the 18O labeling method revealed a decreased γ- and β-ATP turnover in both breast cancer cells, compared to control. Lower β-ATP[18O] turnover indicates decreased adenylate kinase (AK) flux. Aggressive cancer cells had also reduced fluxes through hexokinase (HK) G-6-P[18O], creatine kinase (CK) [CrP[18O], and mitochondrial G-3-P[18O] substrate shuttle. Decreased CK metabolic flux was linked to the downregulation of mitochondrial MTCK1A in breast cancer cells. Despite the decreased overall phosphoryl flux, overexpression of HK2, AK2, and AK6 isoforms within cell compartments could promote aggressive breast cancer growth.

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KW - oxidative phosphorylation

KW - phosphotransfer network

KW - triple-negative breast cancer

KW - γ-and β-ATP phosphoryl turnover

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Stable Isotope Tracing Uncovers Reduced γ/β-ATP Turnover and Metabolic Flux Through Mitochondrial-Linked Phosphotransfer Circuits in Aggressive Breast Cancer Cells

Abstract

Keywords

ASJC Scopus subject areas

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